YE Mei,ZHANG Min,YANG Su-xin,et al.Optimization of Biolistics Transformation of Embryonic Tips of Soybean ［Glycine max (L.) Merrill］ Mature Seeds[J].Soybean Science,2011,30(01):20-23.[doi:10.11861/j.issn.1000-9841.2011.01.0020]
大豆成熟种子胚尖基因枪法转化体系的优化
- Title:
- Optimization of Biolistics Transformation of Embryonic Tips of Soybean ［Glycine max (L.) Merrill］ Mature Seeds
- 文章编号:
- 1000-9841（2011）01-0020-04
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 以山东省主栽大豆品种“荷豆12”成熟种子胚尖为材料，探讨了6-BA和潮霉素浓度等因素对基因枪法遗传转化的影响，并通过报道基因GUS检测了外源基因的表达和整合情况。结果表明：经5 mg·L-1 6-BA诱导16 h，“荷豆12”胚尖的再生丛生芽数目、丛生芽的伸长速率等综合指标最佳；5~15 mg·L-1的潮霉素多步梯度筛选可以提高转化筛选效率。基因枪转化后的胚尖组织和转基因植株后代的叶片中均可以观察到GUS基因的表达；通过分子鉴定证明外源GUS基因插入到转基因后代的基因组中。
- Abstract:
- To establish an efficient transformation system of embryonic tips of soybean mature seeds, effects of 6-BA and hygromycin B concentration were studied during biolistics transformation of soybean ［Glycine max (L.) Merrill cv ‘Hedou12’］. The UidA (GUS) gene was employed as a reporter gene to reveal the foreign gene expression and integration. Both number and elongation length of multiple shoot reached the highest values at 5 mg·L-1 6-BA for 16 h induction of embryonic tips of mature seeds. The optimal selection scheme of transformants was a gradient concentration of hygromycin B from 5 mg·L-1 to 15 mg·L-1. The expression of GUS gene was observed in both embryonic tips after particle bombardment and young leaves of progeny transformants. The integration of GUS gene into the soybean genome was demonstrated by PCR and sequencing analysis.
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备注/Memo
基金项目：国家转基因重大专项资助项目（2009ZX08009-026B）；山东省自然科学杰出青年基金资助项目（JQ200909）。